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异育银鲫过氧化物还原酶基因的克隆及其表达分析
cDNA Cloning, Characterization and Expression Analysis of Peroxiredoxin Gene in Carassius auratus gibelio
税典章#, 陈胜峰#, 吴萍*, 叶元土, 陈佳, 张晶晶
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作者单位:苏州大学基础医学与生物科学学院, 江苏苏州 215123
中文关键字:异育银鲫;过氧化物还原酶;基因克隆;序列分析;组织分布
英文关键字:Carassius auratus gibelio; peroxiredoxin; cloning; sequence analysis; tissue distribution
中文摘要:采用反转录PCR和cDNA末端快速扩增技术从异育银鲫Carassius auratus gibelio肝脏中克隆了过氧化物还原酶(Prx)基因的cDNA全长序列。结果显示,异育银鲫过氧化物还原酶(CaPrx)基因cDNA全长1 035 bp,开放阅读框长594 bp,编码197个氨基酸。在氨基酸序列的N端和C端各有一个保守的Cys残基,属于2-半胱氨酸类Prxs家族。多序列比对和系统进化树分析表明,异育银鲫与鲫Carassius auratus、青鱼Mylopharyngodon piceus、斑马鱼Danio rerio、犀角金线鲃Sinocyclocheilus rhinocerous和朝鲜鳑鲏Rhodeus uyekii等鱼类的Prx基因具有很高的同源性。实时荧光定量PCR结果显示,CaPrx基因在异育银鲫体内广泛表达,在血细胞和肝脏中表达量最高,在鳃和头肾的表达量较少。在感染鲤疱疹病毒Ⅱ型的个体中,肝脏和脾脏的CaPrx基因表达量显著下降,表明其抗氧化作用因病毒的感染而受到了一定的破坏。
英文摘要:In this study, the full-length cDNA sequence of peroxiredoxin (Prx) gene was cloned from the liver of Carassius auratus gibelio by reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends methods. The structure and speculated function of Prx gene were analyzed. The full-length of CaPrx was comprised of 1 035 nucleotides with a 3'untranslated region (UTR) of 366 bp, 5'UTR of 75 bp and an open reading frame of 594 bp, encoding 197 amino acids. The predicted molecular mass and estimated isoelectric point were 21.83 kD and 5.93. In addition, there was a conservative Cys in the N-terminal and C-terminal, respectively, and this suggested that the CaPrx is belonging to 2-Cys Prxs family. Sequence comparison revealed that the deduced amino acid sequence of CaPrx was highly homologous to the Prx homologs of Mylopharyngodon piceus (98%) and Rattus norvegicus (81%). Furthermore, quantitative real-time PCR analysis showed that CaPrx could be detected in various tissues. The highest expression level was detected in hemocyte and liver followed by hemocyte, and little expression of this gene was detected in head kidney, gills and brain. Compared with the diseased fish, the levels of CaPrx was decreased significantly in liver, which indicated that the CaPrx might be involved in immune response against cyprinid herpesvirus Ⅱ infection in C. auratus gibelio.
2018,37(1): 35-44 收稿日期:2017-07-27
DOI:10.11984/j.issn.1000-7083.20170233
分类号:Q959.4;Q78
基金项目:江苏省水产三新工程重大项目(D2015-12)
作者简介:税典章(1992-),男,水产养殖专业研究生,E-mail:deanshui@outlook.com;陈胜峰(1991-),男,水产养殖专业研究生,E-mail:csf546737977@163.com
*通讯作者:吴萍,E-mail:suzhou_wuping@163.com
参考文献:
陈静, 李婧慧, 刘训猛, 等. 2014. 江苏鲫鱼养殖产业发展现状与产业亟待解决问题[J]. 科学养鱼, 12:1-2.
陈群. 2007. 氧化应激对动物消化道结构与功能影响的研究[D]. 江苏无锡:江南大学:83-87.
林秀秀, 叶元土, 吴萍, 等. 2016. 鲤疱疹Ⅱ型病毒(CyHV-2)感染对异育银鲫(Carassius auratus gibelio)组织器官的损伤作用[J]. 基因组学与应用生物学, 35(3):587-594.
刘文枝, 曾令兵, 张辉, 等. 2013. 异育银鲫感染鲤疱疹病毒Ⅱ型(CyHV-2)后血液生理生化指标变化的研究[J]. 上海海洋大学学报, 22(6):928-935.
潘庭双, 胡贤江, 蒋业林. 2001. 异育银鲫的人工繁殖及苗种培育技术[J]. 渔业现代化, 4:10-11.
任丽平. 2014. 鱼过氧化物还原酶peroxiredoxin基因的分子克隆与表达分析[D]. 浙江舟山:浙江海洋学院:47-49.
孙晶, 薛壮. 2015. 斑马鱼过氧化物还原酶的生物信息学分析[J]. 生物技术, 25(1):36-41.
孙盛明, 戈贤平, 傅洪拓, 等. 2014. 日本沼虾过氧化物还原酶基因的克隆及其表达分析[J]. 中国水产科学, 21(3):474-483.
王俊相, 李玉萍, 孔令富, 等. 2010. 鱼类免疫系统的研究进展[J]. 四川畜牧兽医, 7:29-31.
徐文斌, 孙晓波. 1993. 异育银鲫的生物学特性及其在我省的增养殖前景[J]. 黑龙江水产, 2:38-39.
袁军法, 李莉娟, 黄建. 2013. Ⅱ型鲤疱疹病毒感染异育银鲫对鱼体血液中氧化应激指标的影响[C]. 海口:中国水产学会鱼病专业委员会2013年学术研讨会论文摘要汇编.
张晗. 2014. 苏北:异育银鲫鳃出血病爆发[J]. 海洋与渔业, 8(8):28-29.
章波, 向渝梅, 白云. 2004. 抗氧化蛋白Peroxiredoxin家族研究进展[J]. 生理科学进展, 35(4):352-355.
Dierick JF, Wenders F, Chainiaux F, et al. 2003. Retrovirally mediated overexpression of peroxiredoxin Ⅵ increases the survival of WI-38 human diploid fibroblasts exposed to cytotoxic doses of tert-butylhydroperoxide and UVB[J]. Biogerontology, 4:125-131.
Dong WR, Xiang LX, Shao JZ. 2007. Cloning and characterisation of two natural killer enhancing factor genes (NKEF-A and NKEF-B) in pufferfish, Tetraodon nigroviridis[J]. Fish & Shellfish Immunology, 22:1-15.
Fatma N, Singh DP, Shinohara T, et al. 2001. Transcriptional regulation of the antioxidant protein 2 gene, a thiol-specific antioxidant, by lens epithelium-derived growth factor to protect cells from oxidative stress[J]. Journal of Biological Chemistry, 276(52):48899-48907.
Fujii J, Ikeda Y. 2002. Advances in our understanding of peroxiredoxin, amultifunctional, mammalian redox protein[J]. Redox Report Communications in Free Radical Research, 7(3):123-130.
Hall A, Karplus A, Pool LB. 2009. Typical 2-Cys peroxiredoxins:structure, mechanisms and functions[J]. The FEBS Journal, 276(9):2469-2477.
Hofman B, Hecht HJ, Flohé L. 2002. Peroxiredoxins[J]. Biological Chemistry, 383:347-364.
Kang SW, Rhee SG, Chang TS, et al. 2005. 2-Cys peroxiredoxin function in intracellular signal transduction:therapeutic implications[J]. Trends in Molecular Medicine, 11(12):571-578.
Kumar V, Kitaeff N, Hampton MB, et al. 2009. Reversible oxidation of mitochondrial peroxiredoxin 3 in mouse heart subjected to ischemia and reperfusion[J]. FEBS Letters, 583(6):997-1000.
Lee TH, Kim SU, Yu SL, et al. 2003. Peroxiredoxin Ⅱ is essential for sustaining life span of erythrocytes in mice[J]. Blood, 101:5033-5038.
Livak KJ, Schmittgen TD. 2001.Analysis of relative gene expression data using real-time quantitative PCR and the 2 (delta delta C(T)) method[J]. Methods, 25:402-408.
Maningas MBB, Koyama T, Kondo H, et al. 2008. A peroxiredoxin from kuruma shrimp, Marsupenaeus japonicus, inhibited by peptidoglycan[J]. Developmental & Comparative Immunology, 32:198-203.
Mu CK, Zhao JM, Wang LL, et al. 2009. Molecular cloning and characterization of peroxiredoxin 6 from Chinese mitten crab Eriocheir sinensis[J]. Fish & Shellfish Immunology, 26:821-827.
Nonn L, Berggren M, Powis G. 2003. Increased expression of mitochondrial peroxiredoxin-3 (thioredoxin peroxidase-2) protects cancer cells against hypoxia and drug-induced hydrogen peroxide-dependent apoptosis[J]. Molecular Cancer Research, 1(9):682-689.
Nordberg J, Arnér ES. 2001. Reactive oxygen species, antioxidants, and the mammalian thioredoxin system[J]. Free Radical Biology & Medicine, 31(11):1287-1312.
Ren L, Xu T, Wang R, et al. 2013. Miiuy croaker (Miichthys miiuy) Peroxiredoxin 2:molecular characterization, genomic structure and immune response against bacterial infection[J]. Fish & Shellfish Immunology, 34(2):556-563.
Schröder E, Brennan JP, Eaton P, et al. 2008. Cardiac peroxiredoxins undergo complex modifications during cardiac oxidant stress[J]. American Journal of Physiology-Heart and Circulatory Physiology, 295:H425-H433.
Singh AK, Shichi H. 1998. A novel glutathione peroxidase in bovine eye. Sequence analysis, mRNA level, and translation[J]. Journal of Biological Chemistry, 273(40):26171-26178.
Taylor DE, Ghio AJ, Piantadosi CA. 1995. Reactive oxygen species produced by liver mitochondria of rats in sepsis[J]. Archives of Biochemistry & Biophysics, 316(1):70-76.
Von HG. 1990. The signal peptide[J]. Journal of Membrane Biology, 115(3):195-201.
Woo HA, Chae HZ, Hwang SC, et al. 2003. Reversing the inactivation of peroxiredoxins caused by cysteine sulfinic acid formation[J]. Science, 300(5619):653-656.
Wu C, Cao J, Cao F, et al. 2016. Molecular cloning, characterization and mRNA expression of six peroxiredoxins from black carp Mylopharyngodon piceus in response to lipopolysaccharide challenge or dietary carbohydrate[J]. Fish & Shellfish Immunology, 50:210-222.
Yeh HY, Klesius PH. 2007. cDNA cloning, characterization, and expression analysis of channel catfish (Ictalurus punctatus Rafinesque, 1818) peroxiredoxin 6 gene[J]. Fish Physiology & Biochemistry, 33(3):233-239.
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